来自同一牙槽裂患者的骨骼部位特异性人骨髓间充质干细胞的血管生成变异:一项比较研究。

The angiogenic variation of skeletal site-specific human BMSCs from same alveolar cleft patients: a comparative study.

作者信息

Du Yifei, Jiang Fei, Liang Yi, Wang Yuli, Zhou Weina, Pan Yongchu, Xue Mingfei, Peng Yan, Yuan Huan, Chen Ning, Jiang Hongbing

机构信息

Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University, Number 136, Hanzhong Road, Nanjing, 210029, People's Republic of China.

Department of Oral and Maxillofacial Surgery, Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, 210029, People's Republic of China.

出版信息

J Mol Histol. 2016 Apr;47(2):153-68. doi: 10.1007/s10735-016-9662-7. Epub 2016 Feb 4.

Abstract

Tissue engineering strategies hold great potential for alveolar cleft reconstruction. Bone marrow stromal cells (BMSCs) from iliac crest and craniofacial regions are candidate seeding cells with site-specific characteristics and bone-repairing properties. Craniofacial BMSCs seem to possess stronger multipotency and osteogenic capabilities than BMSCs isolated from iliac crest. However, the angiogenic capabilities of these two type cell is rarely reported. We obtained human BMSCs (hBMSCs) of maxilla (M-hBMSCs) and iliac crest (I-hBMSCs) from same alveolar cleft patients to investigate the agiogenic variations using co-culture system with human umbilical vein endothelial cells (HUVECs). From in vitro comparison, M-hBMSCs allowed HUVECs to form more tube-like structures and sprouting angiogenesis by tube formation assays and 3D fibrin vasculogenic assay, respectively. By transplantation in vivo, M-hBMSCs enhanced larger size vessel like structures distributed the entire implants compared with I-hBMSCs. Western blotting was used to assess the angiogenesis related factors including hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). The results showed a significant higher expression of bFGF protein in M-hBMSCs and HUVECs co-culture system both in vitro and in vivo. As bFGF could promote migration and proliferation of endothelial cells, scratch wound healing and transwell migration assays were performed as well as MTT assays and cell cycle analysis. The data suggested the effect of M-hBMSCs on HUVECs was stronger than I-hBMSCs. Taken together, these results indicated that craniofacial BMSCs seemed to have greater angiogenesis capability than iliac crest BMSCs and this might be associated with the different levels of bFGF protein expression in co-culture system.

摘要

组织工程策略在牙槽嵴裂修复方面具有巨大潜力。来自髂嵴和颅面部区域的骨髓间充质干细胞(BMSCs)是具有部位特异性特征和骨修复特性的候选种子细胞。颅面部BMSCs似乎比从髂嵴分离的BMSCs具有更强的多能性和成骨能力。然而,这两种类型细胞的血管生成能力鲜有报道。我们从同一牙槽嵴裂患者获取上颌骨的人BMSCs(M-hBMSCs)和髂嵴的人BMSCs(I-hBMSCs),通过与人脐静脉内皮细胞(HUVECs)共培养系统来研究血管生成差异。通过体外比较,M-hBMSCs分别通过管形成试验和三维纤维蛋白血管生成试验使HUVECs形成更多管状结构和发芽血管生成。通过体内移植,与I-hBMSCs相比,M-hBMSCs增强了分布于整个植入物的更大尺寸血管样结构。采用蛋白质印迹法评估血管生成相关因子,包括肝细胞生长因子(HGF)、血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)。结果显示,在体外和体内,M-hBMSCs与HUVECs共培养系统中bFGF蛋白表达均显著更高。由于bFGF可促进内皮细胞迁移和增殖,还进行了划痕伤口愈合试验、Transwell迁移试验以及MTT试验和细胞周期分析。数据表明M-hBMSCs对HUVECs的作用强于I-hBMSCs。综上所述,这些结果表明颅面部BMSCs似乎比髂嵴BMSCs具有更强的血管生成能力,这可能与共培养系统中bFGF蛋白表达水平不同有关。

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